F. Hoffmann-La Roche, Grenzacherstrasse 124, 4070 Basel, Switzerland.
Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 May 15;1116:1-8. doi: 10.1016/j.jchromb.2019.03.030. Epub 2019 Mar 26.
Hydrolysis of the non-ionic surfactant polysorbate upon long-term storage poses significant challenges to development of biopharmaceutical liquid formulations. Low concentrations of intact surfactant may compromise its protective properties and thus affect protein stability. In addition, accumulation of polysorbate hydrolysis products is increasingly put into context with the formation of visible and subvisible particulates based on the low solubility of the main degradation products. Despite of this potential negative impact on product quality, quantification of the released free fatty acids is performed commonly in an indirect and consequently insensitive manner by determining the remaining PS content or by cumbersome methods, which are unsuitable for routine testing in quality control laboratories. For this purpose, this study describes the development and qualification of a label-free, reliable liquid-chromatography single quad mass detector (LC-QDa)-based method capable of resolving slight changes in the free fatty acid profile which can be readily integrated into quality control facilities. The practical utility of the herein described method is outlined by a case study on the real-time storage stability of a formulated monoclonal antibody.
非离子表面活性剂聚山梨酯在长期储存过程中的水解对生物制药液体制剂的开发构成了重大挑战。低浓度的完整表面活性剂可能会破坏其保护性能，从而影响蛋白质稳定性。此外，基于主要降解产物的低溶解度，聚山梨酯水解产物的积累与可见和亚可见颗粒的形成越来越相关。尽管如此，由于释放的游离脂肪酸的定量分析通常是通过间接的、因此不敏感的方法来进行的，即通过测定剩余的 PS 含量或繁琐的方法来进行，这些方法不适合质量控制实验室的常规测试。为此，本研究描述了一种无标记、可靠的基于液相色谱单四级杆质量检测器（LC-QDa）的方法的开发和验证，该方法能够分辨游离脂肪酸谱的细微变化，可方便地整合到质量控制设施中。通过对单克隆抗体配方实时储存稳定性的案例研究，概述了本文所述方法的实际应用。
